Cytotoxic compounds which are inhibitors of the polymerisation of tubulin

ABSTRACT

The invention relates to compounds which are inhibitors of the polymerization of tubulin, to the methods for the production thereof, and to the uses of same.

FIELD OF THE INVENTION

The present invention relates to compounds which are inhibitors of thepolymerisation of tubulin, to methods for the preparation thereof, andto the uses of same.

STATE OF THE ART

Cancer is a major cause of death around the world, leading to 7.6million deaths in 2008, or about 13% of global mortality. According toWHO estimates, mortality due to cancer will continue to increase toexceed 13 million deaths in 2030. About 30% of cancer deaths are due tofive principal behavioural and dietary risk factors: (i) high body-massindex, (ii) low consumption of fruits and vegetables, (iii) lack ofphysical exercise, (iv) addiction to smoking and (v) alcoholconsumption. Cancer can be reduced and checked by applying strategiesfounded on evidence-based prevention, early detection, patient care andspecific treatments. Concerning the latter point, three principalmethods of cancer treatment exist: surgery, radiotherapy andchemotherapy. For certain types of cancers, it is also possible to callupon hormone therapy, immunotherapy or recent targeted therapies. As thecase may be, these methods may be prescribed alone, successively or incombination. Chemotherapy or polychemotherapy consists in using severalcompounds which are cytotoxic to cancer cells. Unlike surgery, they aresystemic treatments which concern the whole body, not a particularorgan, and which will however have repercussions on healthy cells suchas side effects of varying severity.

One characteristic of cancer cells is to divide very rapidly to enablethe tumour to develop. This characteristic is “targeted” by variousagents which disrupt cell division, such as, for example, DNAintercalators (daunorubicin, doxorubicin, mitoxandrone, etc.),inhibitors of topoisomerases I and II which modify the tertiarystructure of DNA (irinotecan, etoposide, etc.), and even mitotic spindlepoisons.

The importance of microtubules as anti-cancer targets is underlined bythe clinical use of several microtubule-targeting agents in cancertreatment, such as Vinca-type alkaloids of the Madagascar periwinkle andtaxanes. Periwinkle alkaloids (vincristine, vinblastine, etc.) establisha specific bond to tubulin and inhibit its polymerisation intomicrotubules, thus preventing mitotic spindle formation. In turn,taxanes (paclitaxel, docetaxel) prevent tubulin depolymerization andalso disrupt tubulin-microtubule equilibrium. Thus, mitotic spindledisassembly is blocked, which stops mitosis and leads to cell death.Docetaxel is recommended in the treatment of a large number of cancers.It is used alone or in combination with an anti-inflammatory(prednisone, etc.) in advanced prostate cancer (the leading cancer inmen in France). In the case of breast cancers (the leading cancer inwomen), docetaxel is used alone or in combination with other anti-tumouragents (doxorubicin, trastuzumab, etc.). In non-small-cell lung cancer(the second-leading cancer in men and the third in women) it has shownits efficacy alone or combined with cis-platinum. Lastly, docetaxel, incombination with 5-fluorouracil, is given to treat cases of stomach,head and neck cancers. Taxol has found application in first-linetreatments of breast cancer in combination with an anthracycline and ofovarian cancer in combination with cis-platinum.

Despite the therapeutic efficacy of these treatments, these variouscompounds have many side effects (alopecia, neutropenia, nausea,mucositis, muscle pain, etc.). For example, periwinkle alkaloids havebeen associated with problems of neuro-, haemato- and cardiotoxicity.Furthermore, resistance phenomena due to phenotypic changes of cancercells are increasingly observed, such as, for example, in the case ofhormone-independent prostate cancers treated with docetaxel.

Research has been undertaken to identify and develop novel moleculeswhich disrupt the assembly of tubulin into microtubules.

In this context, combretastatin A-4 (CA-4), a member of thecombretastatin family, natural stilbenes isolated by G. R. Pettit fromthe bark of a South-African willow tree, Combretum caffrum ¹, has beenidentified. CA-4 is a stilbene in the Z-configuration substituted withtwo aromatic nuclei of type 3,4,5-trimethoxyphenyl (ring A) and3-hydroxy-4-methoxyphenyl (ring B). CA-4 proved to be highly cytotoxic(IC₅₀=1-2 nM) to many human cancer lines as well as to lines resistantto conventional therapies using, for example, daunorubicin². CA-4 isalso a mitotic spindle poison since it strongly inhibits tubulinassembly into microtubules by binding to the colchicine site (IC₅₀=1μM). It has also been shown that CA-4 exhibits a nanomolaranti-vascular³ activity in vitro by inhibition of endothelial cellproliferation. A water-soluble prodrug of CA-4, fosbretabulin (CA-4P),is currently under phase III clinical development for treatment ofthyroid cancer and under phase II clinical development for treatments ofnon-small-cell lung cancer and for treatments of cis-platinum-resistantovarian cancer. Similarly, the hydrochloride of a structural analogue ofCA-4, ombrabulin, is currently used to treat advanced soft tissuesarcomas.

While these compounds prove to be effective, they have a chemicalinstability attributable to isomerization of the ethylenic double bond.

Isocombretastatin A-4 (CA-4)⁴, an unnatural isomer of CA-4, whosebiological profile (cytotoxicity, tubulin polymerisation inhibition,induction of apoptosis, etc.) is absolutely identical to that of thenatural molecule, without however exhibiting the risk of isomerization,was recently identified. This molecule, which is particularly stable, ismetabolized only very little (<10%) in the presence of hepatocytes.⁵

It has also been shown that it is possible to reduce the double bond ofisoCA-4 without a substantial loss of anti-tumour activity.⁶ Forexample, isoerianin, an unnatural isomer of erianin, proved to becytotoxic at nanomolar concentrations (25<IG₅₀<45 nM) on many humantumour lines and inhibits tubulin polymerisation at micromolarconcentrations by blocking the cell cycle at the G2/M phase.

The combretastatin A-4 analogues synthesized to date have a3,4,5-trimethophenyl moiety. As this group is suspected of being thecause of side effects such as neurotoxicity or cardiotoxicity, a needexists to have available novel agents which inhibit the polymerisationof tubulin. Ideally, these compounds will have to: (i) be effective atdoses which exhibit little or no toxicity in man, (ii) be stable andeasy to produce industrially, (iii) be water-soluble to simplify theirmode of administration, (iv) have an identified mechanism of action and,(v) be free, if possible, of side effects.

BRIEF DESCRIPTION OF THE INVENTION

The present invention relates to a compound of the following formula(I):

wherein:

-   -   R₂ and R₃ are different and one of the two represents a group A₁        of the following formula:

-   -   -   wherein:            -   the dashed bond is present or absent, preferably                present;            -   Z₁ and Z₂ represent, independently of each other, a                hydrogen atom, a halogen atom or a methyl group;            -   (Het)Aryl represents an aryl or heteroaryl group, said                heteroaryl being selected from indolyl, benzothiophenyl                and benzofuranyl groups, wherein said aryl or heteroaryl                may be substituted with one or more substituents                selected from a halogen atom, a hydroxyl group, —OMe,                —SMe, —OCX₃ with X indicating a halogen atom, —NH₂,

with R indicating a C₁-C₆ alkyl group,

with m representing 1 or 2,

with m representing 1 or 2, and

-   -   the other then representing:    -   a hydrogen atom;    -   a halogen atom;    -   a hydroxyl group;    -   a cyano group;    -   a —COYR′ group with Y indicating O or N and R′ indicating H or a        C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group;    -   a —SO₂NR′R″ group with R′, R″ each indicating, independently of        each other, H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a        C₂-C₄ alkynyl group, an aryl group, a heteroaryl group;    -   a —NHSO₂R′ group with R′ indicating a C₁-C₆ alkyl group, a C₂-C₄        alkenyl group, a C₂-C₄ alkynyl group, an aryl group, a        heteroaryl group;    -   a C₁-C₆ alkyl group;    -   a C₂-C₄ alkenyl group;    -   a C₂-C₄ alkynyl group;    -   a C₁-C₆ alkoxy group;    -   or a —NR′R″ group with R′ and R″ representing, independently of        each other, a hydrogen or an alkyl group;    -   A and E each represent a hydrogen atom or A and E take part in        the structure of a fused aromatic ring of the following formula:

-   -   wherein A, B and E represent, independently of each other, a        carbon or nitrogen atom, n represents 0 or 1 and R₄, R₅, R₆, R₇        are as described below;    -   X, Y and Z represent, independently of each other, a carbon or        nitrogen atom provided that if X and Z represent a nitrogen        atom, Y represents a carbon atom;    -   at least one of A, B, E, X, Y and Z represents a nitrogen atom;    -   R₁, R₄, R₅, R₆, R₇, if present, represent, independently of each        other, a hydrogen atom, a halogen atom, a hydroxyl group, a        C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group,        a C₁-C₆ alkoxy group or a —NR′R″ group with R′ and R″        representing, independently of each other, a hydrogen or a C₁-C₆        alkyl group;        as well as the pharmaceutically acceptable salts, the        stereoisomers and the prodrugs thereof.

The present invention also relates to a pharmaceutical compositioncomprising at least one compound of the present invention.

The present invention also relates to compounds or compositions of thepresent invention for use as a medicine, in particular for treating orpreventing proliferative diseases or as an anti-vascular medicine.

DEFINITIONS

The term “halogen”, as used in the description of the present invention,refers to fluorine, chlorine, bromine and iodine atoms. Advantageously,it will be fluorine, bromine and chlorine and more advantageouslyfluorine or chlorine.

The term “C₁-C₆ alkyl”, as used in the description of the presentinvention, refers to any saturated hydrocarbon group comprising 1 to 6carbon atoms, linear or branched, particularly methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexylgroups.

The term “C₂-C₄ alkenyl”, as used in the description of the presentinvention, refers to any hydrocarbon group comprising 2 to 4 carbonatoms, linear or branched, and comprising at least one double bond, suchas a vinyl (ethenyl) group.

The term “C₂-C₄ alkynyl”, as used in the description of the presentinvention, refers to any hydrocarbon group comprising 2 to 4 carbonatoms, linear or branched, and comprising at least one triple bond, suchas an ethynyl or propynyl group.

The term “C₁-C₆ alkoxy”, as used in the description of the presentinvention, refers to any —O-alkyl group, the alkyl being as definedabove. Examples of alkoxy groups include methoxy, ethoxy, propoxy,n-butoxy, iso-butoxy and tert-butoxy groups.

The term “(Het)Aryl”, as used in the description of the presentinvention, refers to an aryl or heteroaryl.

The term “aryl”, as used in the description of the present invention,refers to one or more aromatic rings having 5 to 10 carbon atoms, whichmay be fused. In particular, aryl groups may be monocyclic or bicyclicgroups, such as for example phenyl or naphthyl groups. Advantageously,the aryl group is a phenyl.

The term “heteroaryl”, as used in the description of the presentinvention, refers to an aromatic group comprising 5 to 10 cyclic atoms.Cyclic atoms comprise carbon atoms and one or more heteroatoms, such asfor example sulphur, nitrogen or oxygen atoms. The heteroaryl accordingto the present invention may consist of one or two fused rings.Preferably, the heteroaryl group will be an indolyl, benzothiophenyl,benzofuranyl or benzoimidazolyl group.

The term “cyano”, as used in the description of the present invention,refers to a —CN group.

The formula “—COYR”, as used in the description of the presentinvention, refers to an acid or an ester when Y is O, an amide when Y isN.

The formula “—SO₂NR′R″” or “—NHSO₂R′”, as used in the description of thepresent invention, refers to a sulphonamide.

The expression “pharmaceutically acceptable”, as used in the descriptionof the present invention, refers to that which is useful in thepreparation of a pharmaceutical composition, which is generally safe,nontoxic and neither biologically nor otherwise undesirable and which isacceptable for veterinary and/or human pharmaceutical use.

The expression “pharmaceutically acceptable salts”, as used in thedescription of the present invention, refers to salts of a compoundwhich are pharmaceutically acceptable, as defined herein, and which havethe desired pharmacological activity of the parent compound. Such saltsinclude:

(1) hydrates and solvates,

(2) acid addition salts formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulphuric acid, nitric acid, phosphoric acid andthe like; or formed with organic acids such as acetic acid,benzenesulfonic acid, benzoic acid, camphosulfonic acid, citric acid,ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid,glutamic acid, glycolic acid, hydroxynaphthoic acid,2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid,mandelic acid, methanesulfonic acid, muconic acid, 2-naphthalenesulfonicacid, propionic acid, salicylic acid, succinic acid,dibenzoyl-L-tartaric acid, tartaric acid, p-toluenesulfonic acid,trimethylacetic acid, trifluoroacetic acid and the like. Advantageously,it is hydrochloric acid; or

(3) salts formed when an acidic proton present in the parent compoundeither is replaced with a metal ion, for example an alkaline metal ion,an alkaline-earth metal ion; or coordinates with an organic or inorganicbase. Acceptable organic bases include diethanolamine, ethanolamine,N-methylglucamine, triethanolamine, tromethamine and the like.Acceptable inorganic bases include aluminium hydroxide, calciumhydroxide, potassium hydroxide, sodium carbonate and sodium hydroxide.Advantageously, the acidic proton is displaced by a Na⁺ ion,particularly using sodium hydroxide. Acid addition salts are formedparticularly with an amine functional group or a pyridine. Base additionsalts are formed particularly with a carboxylic acid (—COOH), phosphate(—OP(O)(OH)₂) or sulphate (—OSO₃H) functional group.

The term “stereoisomers”, as used in the description of the presentinvention, refers to diastereoisomers or enantiomers. They are thusconfigurational isomers. Stereoisomers which are not mirror images ofone another are thus referred to as “diastereoisomers”, andstereoisomers which are mirror images of one another but arenon-superimposable are referred to as “enantiomers”, also called“optical isomers”. A carbon atom linked to four non-identicalsubstituents is called a “chiral centre”. When a molecule has such achiral centre, it is called chiral and has two enantiomer forms. When amolecule has several chiral centres, then it will have severaldiastereoisomer and enantiomer forms. An equimolar mixture of twoenantiomers is called a racemic mixture.

The term “prodrug”, as used in the description of the present invention,refers to a compound which is administered in an inactive (or lessactive) form and which is metabolized in vivo, particularly by theaction of enzymes or gastric acid, in an active (or more active) form.The use of a prodrug improves in particular the physicochemicalparameters of a molecule, such as solubility, as well as thepharmacokinetics (vectorization, bioavailability, etc.), in order topromote its assimilation by an organism after administration. Inparticular, a prodrug of a molecule bearing an amino group (NH₂) canresult particularly from the acylation or the phosphorylation of thisamino group. When a molecule bears a hydroxy group (OH), the prodrug canresult particularly from the acylation or the phosphorylation of thishydroxy group.

The expression “compounds of the present invention” as used in thepresent description refers to compounds of formula (I), (I′), (Ib) or(Ic) as defined in a detailed manner below.

DETAILED DESCRIPTION OF THE INVENTION

The inventors have shown that compounds of the present invention disruptthe assembly of tubulin into microtubules. These compounds havenanomolar cytotoxic activities on various human cancer lines, includinglines resistant to common treatments. These compounds inhibit thepolymerisation of tubulin at micromolar concentrations, evensub-micromolar concentrations, thus constituting therapeutic compoundsof choice.

Due to their chemical structure, compounds of the present invention havea low risk of being metabolized, unlike CA-4 derivatives which aresubject to (i) demethylation reactions on the 3,4,5-trimethoxyphenylring or (ii) isomerization of the double bond leading to a substantialloss of activity.

Compounds of the Present Invention

The invention thus has as an object compounds of the following formula(I):

wherein:

-   -   R₂ and R₃ are different and one of the two represents a group A₁        of the following formula:

-   -   -   wherein:            -   the dashed bond is present or absent, preferably                present;            -   Z₁ and Z₂ represent, independently of each other, a                hydrogen atom, a halogen atom or a methyl group;            -   (Het)Aryl represents an aryl or heteroaryl group, said                heteroaryl being selected from indolyl, benzothiophenyl                and benzofuranyl groups, wherein said aryl or heteroaryl                may be substituted with one or more substituents                selected from a halogen atom, a hydroxyl group, —OMe,                —SMe, —OCX₃ with X indicating a halogen atom, —NH₂,

with R indicating a C₁-C₆ alkyl group,

with m representing 1 or 2,

with m representing 1 or 2, and

-   -   the other representing:        -   a hydrogen atom;        -   a halogen atom;        -   a hydroxyl group;        -   a cyano group;        -   a —COYR′ group with Y indicating O or N and R′ indicating H            or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄            alkynyl group;        -   a —SO₂NR′R″ group with R′, R″ each indicating, independently            of each other, H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl            group, a C₂-C₄ alkynyl group;        -   a —NHSO₂R′ group with R′ indicating a C₁-C₆ alkyl group, a            C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group, an aryl group, a            heteroaryl group;        -   a C₁-C₆ alkyl group;        -   a C₂-C₄ alkenyl group;        -   a C₂-C₄ alkynyl group;        -   a C₁-C₆ alkoxy group; or        -   a —NR′R″ group with R′ and R″ representing, independently of            each other, a hydrogen or an alkyl group;    -   A and E each represent a hydrogen atom or A and E take part in        the structure of a fused aromatic ring of the following formula:

-   -   wherein A, B and E represent, independently of each other, a        carbon or nitrogen atom, n represents 0 or 1 and R₄, R₅, R₆, R₇        are as described below;    -   X, Y and Z represent, independently of each other, a carbon or        nitrogen atom provided that if X and Z represent a nitrogen        atom, Y represents a carbon atom;    -   at least one of A, B, E, X, Y and Z represents a nitrogen atom;    -   R₁, R₄, R₅, R₆, R₇, if present, represent, independently of each        other:        -   a hydrogen atom;        -   a halogen atom;        -   a hydroxyl group;        -   a C₁-C₆ alkyl group;        -   a C₂-C₄ alkenyl group;        -   a C₂-C₄ alkynyl group;        -   a C₁-C₆ alkoxy group; or        -   a —NR′R″ group with R′ and R″ representing, independently of            each other, a hydrogen or a C₁-C₆ alkyl group;            as well as the pharmaceutically acceptable salts, the            stereoisomers and the prodrugs thereof.

Preferably, the invention has as an object compounds of the followingformula (I′):

wherein:

-   -   n represents 0 or 1;    -   A, B and E represent, independently of each other, a carbon or        nitrogen atom;    -   X, Y and Z represent, independently of each other, a carbon or        nitrogen atom provided that if X and Z represent a nitrogen        atom, Y represents a carbon atom;    -   at least one of A, B, E, X, Y and Z represents a nitrogen atom;    -   R₁, R₄, R₅, R₆, R₇, if present, represent, independently of each        other:        -   a hydrogen atom;        -   a halogen atom;        -   a hydroxyl group;        -   a C₁-C₆ alkyl group;        -   a C₂-C₄ alkenyl group;        -   a C₂-C₄ alkynyl group;        -   a C₁-C₆ alkoxy group; or        -   a —NR′R″ group with R′ and R″ representing, independently of            each other, a hydrogen or a C₁-C₆ alkyl group;    -   R₂ and R₃ are different and one of the two represents a group A₁        of the following formula:

-   -   -   wherein:            -   the dashed bond is present or absent, preferably                present;            -   Z₁ and Z₂ represent, independently of each other, a                hydrogen atom, a halogen atom or a methyl group;            -   (Het)Aryl represents an aryl or heteroaryl group, said                heteroaryl being selected from indolyl, benzothiophenyl                and benzofuranyl groups, wherein said aryl or heteroaryl                may be substituted with one or more substituents                selected from a halogen atom, a hydroxyl group, —OMe,                —SMe, —OCX₃ with X indicating a halogen atom, —NH₂,

with R indicating a C₁-C₆ alkyl group,

with m representing 1 or 2,

with m representing 1 or 2, and

-   -   and the other represents:        -   a hydrogen atom;        -   a halogen atom;        -   a hydroxyl group;        -   a cyano group;        -   a —COYR′ group with Y indicating O or N and R′ indicating H            or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄            alkynyl group;        -   a —SO₂NR′R″ group with R′, R″ each indicating, independently            of each other, H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl            group, a C₂-C₄ alkynyl group;        -   a —NHSO₂R′ group with R′ indicating a C₁-C₆ alkyl group, a            C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group, an aryl group, a            heteroaryl group;        -   a C₁-C₆ alkyl group;        -   a C₂-C₄ alkenyl group;        -   a C₂-C₄ alkynyl group;        -   a C₁-C₆ alkoxy group;        -   a —NR′R″ group with R′ and R″ representing, independently of            each other, a hydrogen or a C₁-C₆ alkyl group;            as well as the pharmaceutically acceptable salts, the            stereoisomers and the prodrugs thereof.

The expression “R₁, R₄, R₅, R₆ or R₇, if present”, as used in thedescription of the present invention, means that the R₁, R₄, R₅, R₆ orR₇ groups will be present if the valency of the atom to which the groupis, or would be bound allows it. Persons skilled in the art will be ableeasily to determine if such a group is present.

In certain embodiments, the invention has as an object compounds offormula (I′) as represented above wherein R₃ is a group A₁ asrepresented above, or compounds of the following formula (Ib):

wherein:

-   -   n, A, B, E, X, Y, Z, R₁, R₄, R₅, R₆, R₇, (Het)Aryl, Z₁ and Z₂        are as described above; and    -   R₂ represents:        -   a hydrogen atom;        -   a halogen atom;        -   a hydroxyl group;        -   a cyano group;        -   a —COYR′ group with Y indicating O or N and R′ indicating H            or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄            alkynyl group;        -   a SO₂NR′R″ group with R′, R″ each indicating, independently            of each other, H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl            group, a C₂-C₄ alkynyl group;        -   a —NHSO₂R′ group with R′ indicating a C₁-C₆ alkyl group, a            C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group, an aryl group, a            heteroaryl group;        -   a C₁-C₆ alkyl group;        -   a C₂-C₄ alkenyl group;        -   a C₂-C₄ alkynyl group;        -   a C₁-C₆ alkoxy group;        -   or a —NR′R″ group with R′ and R″ representing, independently            of each other, a hydrogen or an alkyl group.

In certain embodiments, the invention has as an object compounds offormula (I′) or (Ib) wherein the (Het)Aryl group represents a phenylgroup which may be substituted with one or more substituents selectedfrom a halogen atom, a hydroxyl group, —OMe, —SMe, —OCX₃ with Xindicating a halogen atom, —NH₂,

with R indicating a C₁-C₆ alkyl group,

with m representing 1 or 2,

with m representing 1 or 2 and

Preferably, the (Het)Aryl group represents a substituted phenyl grouphaving the following formula:

wherein:

-   -   R₈ represents a hydrogen atom, a halogen atom, a hydroxyl group,        —OMe, —SMe, —NH₂,

with R indicating a C₁-C₆ alkyl group,

with m representing 1 or 2,

with m representing 1 or 2 or

and

-   -   R₉ represents a C₁-C₆ alkoxy group, particularly —OMe, —SMe or        —OCX₃ with X indicating a halogen atom.

Preferably, R₈ represents a hydrogen atom, a halogen atom, preferablyfluorine or chlorine, a hydroxyl group or a —NH₂ group.

Preferably, R₉ represents a C₁-C₆ alkoxy group, preferably C₁-C₄,preferably methoxy, —SMe or —OCX₃ with X indicating a halogen atom,preferably fluorine.

Thus, in certain embodiments, the invention has as an object compoundsof the following formula (Ic):

wherein:

-   -   n, A, B, E, X, Y, Z, R₁, R₄, R₅, R₆, R₇ are as described above;    -   R₂ represents:        -   a hydrogen atom;        -   a halogen atom;        -   a hydroxyl group;        -   a cyano group;        -   a —COYR′ group with Y indicating O or N and R′ indicating H            or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄            alkynyl group;        -   a —SO₂NR′R″ group with R′, R″ each indicating, independently            of each other, H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl            group, a C₂-C₄ alkynyl group;        -   a —NHSO₂R′ group with R′ indicating a C₁-C₆ alkyl group, a            C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group, an aryl group, a            heteroaryl group;        -   a C₁-C₆ alkyl group;        -   a C₂-C₄ alkenyl group;        -   a C₂-C₄ alkynyl group;        -   a C₁-C₆ alkoxy group;        -   or a —NR′R″ group with R′ and R″ representing, independently            of each other, a hydrogen or an alkyl group;    -   the dashed bond is absent or present, preferably present;    -   Z₁ and Z₂ represent, independently of each other, a hydrogen        atom, a halogen atom or a methyl group;    -   R₈ represents a hydrogen atom, a halogen atom, a hydroxyl, —OMe,        —SMe, —NH₂,

with R indicating a C₁-C₆ alkyl group,

with m representing 1 or 2,

with m representing 1 or 2 or

and

-   -   R₉ represents a C₁-C₆ alkoxy group, particularly —OMe, —SMe or        —OCX₃ with X indicating a halogen atom;        as well as the pharmaceutically acceptable salts, the        stereoisomers and the prodrugs thereof.

Preferably, in the formula (Ic) above, R₈ represents a hydrogen atom, ahalogen atom, preferably fluorine or chlorine, a hydroxyl group, a —NH₂group, or

with m representing 1 or 2.

Preferably, in the formula (Ic) above, R₉ represents a C₁-C₆ alkoxygroup, preferably C₁-C₄, preferably methoxy, —SMe or —OCX₃ with Xindicating a halogen atom, preferably fluorine.

In the formulas (I), (I′), (Ib) and (Ic) above, groups R₁, R₄, R₅, R₆and R₇ when present and group R₂ or R₃ when different from A₁ represent,independently of each other, preferably:

-   -   a hydrogen atom;    -   a halogen atom, preferably chlorine or fluorine;    -   a hydroxyl group;    -   a cyano group;    -   a —COYR′ group with Y indicating O or N and R′ indicating H or a        C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group;    -   a —SO₂NR′R″ group with R′, R″ each indicating, independently of        each other, H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a        C₂-C₄ alkynyl group;    -   a —NHSO₂R′ group with R′ indicating a C₁-C₆ alkyl group, a C₂-C₄        alkenyl group, a C₂-C₄ alkynyl group, an aryl group, a        heteroaryl group;    -   a C₁-C₆ alkyl group, preferably a C₁-C₄ alkyl group, preferably        a methyl;    -   a C₁-C₆ alkoxy group, preferably a C₁-C₄ alkoxy group,        preferably the methoxy group; or    -   a —NR′R″ group with R′ and R″ representing, independently of        each other, a hydrogen or a C₁-C₆ alkyl group, preferably C₁-C₄,        preferably —NH₂ or —N(CH₃)₂.

Preferably, compounds of the present invention have the formula (I),(I′), (Ib) or (Ic) wherein Z₁ and Z₂ are identical and represent ahydrogen or fluorine atom.

Preferably, the dashed bond is present.

In certain embodiments, compounds of the present invention have theformula (I′) represented above wherein:

-   -   n, A, B, E, X, Y, Z, R₁, R₄, R₅, R₆, R₇, are as described above;        and    -   R₂ or R₃ represents a group A₂ of the following formula:

wherein:

-   -   the dashed bond is present or absent, preferably present;    -   Z₁ and Z₂ represent, independently of each other, a hydrogen        atom, a halogen atom or a methyl group.

In certain embodiments, compounds of the present invention have theformula (I′) represented above wherein:

-   -   n, A, B, E, X, Y, Z, R₁, R₄, R₅, R₆, R₇, are as described above;        and    -   R₂ or R₃ represent a group A₃ of the following formula:

wherein:

-   -   the dashed bond is present or absent, preferably present;    -   Z₁ and Z₂ represent, independently of each other, a hydrogen        atom, a halogen atom or a methyl group;    -   R₁₀ represents a hydrogen atom or a methyl group.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   X is a nitrogen atom;    -   Z is a carbon atom or a nitrogen atom;    -   A and E are, independently of each other, a nitrogen or carbon        atom; and    -   Y and B are a carbon atom.

In these embodiments, preferably, R₅ is a hydrogen atom and R₇, ifpresent, is a hydrogen atom.

In particular, compounds of the present invention may have the formula(I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1, X and Z are a nitrogen atom and A, E, Y and B        are a carbon atom;    -   n is equal to 1, X is a nitrogen atom and A, E, Z, Y and B are a        carbon atom;    -   n is equal to 1, Z is a nitrogen atom and A, E, X, Y and B are a        carbon atom;    -   n is equal to 1, X, Z and E are a nitrogen atom and A, Y and B        are a carbon atom; or    -   n is equal to 1, X, Z and A are a nitrogen atom and E, Y and B        are a carbon atom.

In particular, compounds of the present invention may have the formula(I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1, X and Z are a nitrogen atom and A, E, Y and B        are a carbon atom;    -   n is equal to 1, X is a nitrogen atom and A, E, Z, Y and B are a        carbon atom

Preferably, when n is equal to 1, X and Z are a nitrogen atom and A, E,Y and B are a carbon atom, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein R₄, R₅, R₆ and R₇represent a hydrogen atom. In certain embodiments, compounds of thepresent invention have the formula (Ib) or (Ic) wherein n is equal to 1,X and Z are a nitrogen atom, A, E, Y and B are a carbon atom, R₄, R₅, R₆and R₇ represent a hydrogen atom and R₂ is as defined above. Preferably,R₂ is different from a hydrogen atom. In other embodiments, compounds ofthe present invention have the formula (I′) wherein n is equal to 1, Xand Z are a nitrogen atom, A, E, Y and B are a carbon atom, R₄, R₅, R₆and R₇ represent a hydrogen atom, R₂ is a group A₁ as defined above andR₃ is as defined above, preferably R₃ is different from a hydrogen atom.

Preferably, when n is equal to 1, X is a nitrogen atom and A, E, Z, Yand B are a carbon atom, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein R₄, R₅, R₆ and R₇represent a hydrogen atom. In certain embodiments, compounds of thepresent invention have the formula (Ib) or (Ic) wherein n is equal to 1,X is a nitrogen atom, A, E, Z, Y and B are a carbon atom, R₄, R₅, R₆ andR₇ represent a hydrogen atom and R₂ is as defined above. Preferably, R₂is different from a hydrogen atom. In other embodiments, compounds ofthe present invention have the formula (I′) wherein n is equal to 1, Xis a nitrogen atom, A, E, Z, Y and B are a carbon atom, R₄, R₅, R₆ andR₇ represent a hydrogen atom, R₂ is a group A₁ as defined above and R₃is as defined above, preferably R₃ is different from a hydrogen atom.

Preferably, when n is equal to 1, X, Z and E are a nitrogen atom and A,Y and B are a carbon atom, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein R₄, R₅ and R₆represent a hydrogen atom. In certain embodiments, compounds of thepresent invention have the formula (Ib) or (Ic) wherein n is equal to 1,X, Z and E are a nitrogen atom, A, Y and B are a carbon atom, R₄, R₅ andR₆ represent a hydrogen atom and R₂ is as defined above. Preferably, R₂is different from a hydrogen atom. In other embodiments, compounds ofthe present invention have the formula (I) wherein n is equal to 1, X, Zand E are a nitrogen atom, A, Y and B are a carbon atom, R₄, R₅ and R₆represent a hydrogen atom, R₂ is a group A₁ as defined above and R₃ isas defined above, preferably R₃ is different from a hydrogen atom.

Preferably, when n is equal to 1, X, Z and A are a nitrogen atom and E,Y and B are a carbon atom, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein R₅, R₆ and R₇ are ahydrogen atom.

In certain embodiments, compounds of the present invention have theformula (Ib) or (Ic) wherein n is equal to 1, X, Z and A are a nitrogenatom and E, Y and B are a carbon atom, R₅, R₆ and R₇ are a hydrogen atomand R₂ is as defined above. Preferably, R₂ is different from a hydrogenatom. In other embodiments, compounds of the present invention have theformula (I′) wherein n is equal to 1, X, Z and A are a nitrogen atom andE, Y and B are a carbon atom, R₅, R₆ and R₇ is a hydrogen atom, R₂ is agroup A₁ as defined above and R₃ is as defined above, preferably R₃ isdifferent from a hydrogen atom.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 0;    -   A and X are nitrogen atoms;    -   E and Z are, independently of each other, a nitrogen or carbon        atom; and    -   Y is a carbon atom.

In these embodiments, preferably, R₅ is a hydrogen atom and R₇, ifpresent, is a hydrogen atom.

In particular, compounds of the present invention may have the formula(I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 0, A, X, E and Z are a nitrogen atom and Y is a        carbon atom;    -   n is equal to 0, A, X and Z are a nitrogen atom and E and Y are        a carbon atom; or    -   n is equal to 0, A and X are a nitrogen atom and E, Y and Z are        a carbon atom.

Preferably, when n is equal to 0, A, X, E and Z are a nitrogen atom andY is a carbon atom, compounds of the present invention have the formula(I′), (Ib) or (Ic) represented above wherein R₅ is a hydrogen atom andR₄ and R₂ are as defined above, preferably R₄ and R₂ are different froma hydrogen atom.

Preferably, when n is equal to 0, A, X and Z are a nitrogen atom and Eand Y are a carbon atom, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein R₂, R₅ and R₇ are ahydrogen atom and R₄ is as defined above, preferably R₄ is differentfrom a hydrogen atom.

Preferably, when n is equal to 0, A and X are a nitrogen atom and E, Yand Z are a carbon atom, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein R₂, R₅ and R₇ are ahydrogen atom and R₄ is as defined above, preferably R₄ is differentfrom a hydrogen atom.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   A, B, E, Y and Z are a carbon atom;    -   X is a nitrogen atom.

In these embodiments, preferably, R₄, R₅, R₆ and R₇ represent a hydrogenatom.

In particular, compounds of the present invention may have the formula(Ib) or (Ic) represented above wherein n is equal to 1, A, B, E, Y and Zare a carbon atom, X is a nitrogen atom and R₂ is as defined above,preferably R₂ is different from a hydrogen atom.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   A, B, E, Y and X are a carbon atom;    -   Z is a nitrogen atom.

In these embodiments, preferably, R₄, R₅, R₆ and R₇ represent a hydrogenatom.

In particular, compounds of the present invention may have the formula(Ib) or (Ic) represented above wherein n is equal to 1, A, B, E, Y and Xare a carbon atom, Z is a nitrogen atom and R₂ is as defined above,preferably R₂ is different from a hydrogen atom.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   A, B, E and X are a carbon atom;    -   Y and Z are a nitrogen atom.

In these embodiments, preferably, R₄, R₅, R₆ and R₇ represent a hydrogenatom.

In particular, compounds of the present invention may have the formula(Ib) or (Ic) represented above wherein n is equal to 1, A, B, E and Xare a carbon atom, Y and Z are a nitrogen atom and R₁ is as definedabove, preferably R₁ is different from a hydrogen atom.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   X, Y, Z and E are a carbon atom;    -   A and B are a nitrogen atom.

In these embodiments, preferably, R₁, R₂ and R₇ represent a hydrogenatom.

In particular, compounds of the present invention may have the formula(Ib) or (Ic) represented above wherein n is equal to 1, X, Y, Z and Eare a carbon atom, A and B are a nitrogen atom and R₅ is as definedabove, preferably R₅ is different from a hydrogen atom.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   A, B and E represent a carbon atom; and    -   X, Y, Z, R₁, R₂, R₃, R₄, R₅, R₆ and R₇ are as described above.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   A, B and E represent a carbon atom;    -   R₄, R₅, R₆ and R₇ represent a hydrogen; and    -   X, Y, Z, R₁, R₂ and R₃ are as described above.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   A, B and E represent a carbon atom;    -   R₄, R₅, R₆ and R₇ represent a hydrogen;    -   X and Y represent a nitrogen atom;    -   Z represents a carbon atom; and    -   R₁, R₂ and R₃ are as described above.

In certain embodiments, compounds of the present invention have theformula (I′), (Ib) or (Ic) represented above wherein:

-   -   n is equal to 1;    -   A, B and E represent a carbon atom;    -   R₄, R₅, R₆ and R₇ represent a hydrogen;    -   X and Y represent a nitrogen atom;    -   Z represents a carbon atom;    -   R₁ is absent;    -   R₂ represents a hydrogen, a halogen, a C₁-C₆ alkyl group, a        C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group, a C₁-C₆ alkoxy group        or a —NR′R″ group with R′ and R″ representing, independently of        each other, a hydrogen or an alkyl group; and    -   R₃ is as described above.

In certain particular embodiments, compounds of the present inventionhave one of the following formulas:

wherein, in an appropriate manner, Z₁, Z₂, R₁, R₂, R₃, R₄, R₅, R₈ and R₉are as defined above and the dashed bond is absent or present.Preferably, the dashed bond is present.

In particular, in certain embodiments, Z₁ and Z₂ each represent ahydrogen or each a fluorine atom and R₁, R₂, R₃, R₄, R₅, R₈ and R₉ areas defined above.

In other embodiments, R₉ indicates a methoxy and Z₁, Z₂, R₁, R₂, R₃, R₄,R₅ and R₈ are as defined above.

In other embodiments, Z₁ and Z₂ each represent a hydrogen or each afluorine atom, R₉ represents a methoxy and R₁, R₂, R₃, R₄, R₅ and R₈ areas defined above.

In particular, compounds of the invention may be selected from:

The absence of a double ethylene bond in compounds of formula (I) solvesthe isomerization problem likely to occur in vivo, leading to drops in(or absences of) cytotoxic activity as is the case, for example, ofCA-4. Reduction of the double bond located between the two aromaticnuclei yields derivatives which are less cytotoxic than theirprecursors.

Method for Synthesizing Compounds of the Present Invention

The invention also has as an object the methods for synthesizingcompounds of the present invention. The synthesis methods are short,advantageously comprising 2 to 3 steps. These methods are compatiblewith industrial requirements.

Coupling an easily accessible tosylhydrazone with a halogenatedderivative bearing the nitrogenous bicycle gives compounds according tothe invention with excellent yields, advantageously without having toresort to protection-deprotection steps.

Compounds according to the invention may be prepared according tomethods known to persons skilled in the art, starting withcommercially-available products or prepared according to methods knownto persons skilled in the art.

In particular, compounds of formula (I), (I′), (Ib) and (Ic) may beprepared by a method comprising the following successive steps:

-   -   a) reaction of a compound of formula (II)

-   -   with tosylhydrazine to yield the tosylhydrazone compound of        formula (III)

-   -   b) coupling of the compound of formula (III) with the compound        of formula (IV), (V), (VI) or (VII)

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, X, Y, Z, A, B, E and n are aspreviously defined and X₁ represents a halogen atom ortrifluoromethanesulfonate;

-   -   b) if need be, reduction of compound (I), wherein the dashed        double bond is present, obtained following step b) to yield        compound (I) wherein the dashed double bond is absent;    -   c) separation from the reaction medium of compound (I) obtained        following step b) or c).

Persons skilled in the art will protect, in a suitable manner, if needbe, groups R₁ to R₇ so that only group X₁ reacts or adapt as aconsequence the synthesis strategy.

For example, when groups R₁, R₂, R₃, R₄, R₅, R₆ and/or R₇ (if present)represent a halogen atom:

-   -   X₁ is an iodine atom if groups R₁, R₂, R₃, R₄, R₅, R₆ and R₇ (if        present) are a bromine, chlorine or fluorine atom;    -   if groups R₁, R₂, R₃, R₄, R₅, R₆ and R₇ (if present) are a        chlorine or fluorine atom, then X₁ is a bromine atom.

In particular, compounds of formula (I′), (Ib) or (Ic) wherein n isequal to 1, A, B, E and Y are carbon atoms and X and Z are nitrogenatoms may be prepared by a method comprising the following successivesteps:

-   -   a) reaction of a compound of formula (II)

with tosylhydrazine to yield the tosylhydrazone compound of formula(III)

-   -   b) coupling of the compound of formula (III) with the compound        of formula (IV′) or (V′)

-   -   wherein R₂, R₃, R₄, R₅, R₆ and R₇ are as previously defined and        X₁ represents a halogen atom or trifluoromethanesulfonate;    -   c) if need be, reduction of compound (I), wherein the dashed        double bond is present, obtained following step b) to yield        compound (I) wherein the dashed double bond is absent;    -   d) separation from the reaction medium of compound (I) obtained        following step b) or c).

This step c) may be followed by possible additional conventional stepsfor modifying the substituents of the (Het)Aryl group and possibly ofZ₁/Z₂.

The compound thus obtained may be separated from the reaction medium bymethods well-known to persons skilled in the art, such as for example byextraction, solvent evaporation or by precipitation and filtration.

The compound may be further purified if need be by techniques well-knownto persons skilled in the art, such as by recrystallization if thecompound is crystalline, by distillation, by column chromatography onsilica gel or by high-performance liquid chromatography (HPLC).

During step b), the coupling is advantageously carried out in thepresence of a palladium complex, a ligand, in a catalytic quantity, anda base. In particular, the coupling may be carried out by means of apalladium complex such as PdCl₂(MeCN)₂, the ligand dppf in catalyticquantity and the base tBuOLi. In particular, the coupling may be carriedout in the presence of 10% molar PdCl₂(MeCN)₂, 20% molar dppf, 2.2 molarequivalent tBuOLi in the presence of dioxane, at 90° C., for 2 to 4hours.

During step c), the reduction is achieved advantageously byhydrogenation. The hydrogenation is advantageously carried out underhydrogen atmosphere, particularly in the presence of palladium on carbon(Pd/C) as catalyst or possibly PtO₂. Advantageously, 5 to 30 mol %,preferably about 10 mol % catalyst is used during this reaction.Moreover, ethyl acetate will advantageously be used as solvent duringthis step.In particular, compounds of formula (I), (I′), (Ib) and (Ic) may beprepared by a method comprising the following successive steps:

In particular, certain compounds of the present invention may beprepared according to the following scheme:

Uses of Compounds of the Present Invention

In certain aspects, the invention relates to compounds of formula (I),(Ib) or (Ic), as well as to the pharmaceutically acceptable salts, thestereoisomers and the prodrugs thereof, for use as a medicine.

In particular, compounds of the present invention may be used asmedicines which inhibit the polymerisation of tubulin, advantageously asmedicines for treating or preventing proliferative diseases, such ascancer, psoriasis or fibrosis, particularly cancer. In particular,compounds of the invention may be useful in the treatment of a cancer,such as those able to be treated with CA-4 or with taxotere.

Compounds of the present invention may also be used as anti-vascularmedicines.

The invention also relates to a pharmaceutical composition comprising atleast one compound according to the present invention, typically incombination with one or more pharmaceutically acceptable excipients.

In certain aspects, the invention also relates to a pharmaceuticalcomposition comprising at least one compound according to the presentinvention in combination with at least one other active ingredient,particularly an anti-cancer compound, cytotoxic or not, and one or morepharmaceutically acceptable excipients.

In a non-limiting manner, the active ingredients able to be combinedwith the compound of the present invention may be selected from6-mercaptopurine, fludarabine, cladribine, pentostatin, cytarabine,5-fluorouracil, gemcitabine, methotrexate, raltitrexed, irinotecan,topotecan, etoposide, daunorubicin, doxorubicin, epirubicin, idarubicin,pirarubicin, mitoxantrone, chlormethine, cyclophosphamide, ifosfamide,melphalan, chlorambucil, busulfan, carmustine, fotemustine,streptozocin, carboplatin, cisplatin, oxaliplatin, procarbazine,dacarbazine, bleomycin, vinblastine, vincristine, vindesine,vinorelbine, paclitaxel, docetaxel, L-asparaginase, flutamide,nilutamide, bicalutamide, cyproterone acetate, triptorelin, leuprorelin,goserelin, buserelin, formestane, aminoglutethimide, anastrazole,letrozole, tamoxifen, octreotide, lanreotide,(Z)-3-[2,4-dimethyl-5-(2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-1H-pyrrol-3-yl]-propionicacid,4-((9-chloro-7-(2,6-difluorophenyl)-5H-pyrimidol(5,4-d)(2)benzazepin-2-yl)amino)benzoicacid, 5,6-dimethylxanthenone-4-acetic acid and3-(4-(1,2-diphenylbut-1-enyl)phenyl)acrylic acid.

The invention also concerns a pharmaceutical composition comprising atleast one compound according to the present invention in combinationwith an antibody. The composition may comprise one or morepharmaceutically acceptable excipients. The antibody is used to targetthe tumour. In particular, the pharmaceutical composition may compriseat least one compound according to the present invention in combinationwith a monoclonal antibody. The combination of the compound of thepresent invention with the antibody may be achieved in the form of“antibody-compound of the present invention” conjugates. The antibodyand the compound of the present invention are typically bound covalentlyby means of a bond. Persons skilled in the art will be able to determinethe nature of the bond suited to bonding the compound of the presentinvention to an antibody. Thus, in one aspect, the present inventionrelates to a conjugate comprising a compound according to the presentinvention bound covalently to an antibody.

Compounds and compositions according to the invention may beadministered orally, sublingually, parenterally, subcutaneously,intramuscularly, intravenously, transdermally, locally or rectally.

Compounds according to the present invention may be used at dosesbetween 0.01 mg and 1000 mg per day, given in a single dose once per dayor preferably administered in several doses throughout the day, forexample twice per day in equal doses. The dose administered per day isadvantageously between 5 mg and 500 mg, even more advantageously between10 mg and 200 mg. It may be necessary to use doses outside these ranges,which persons skilled in the art will be able to determine.

Compounds according to the invention may be used to decrease or inhibitthe polymerisation of tubulin, particularly in vitro and also in vivo.

The present invention also relates to a pharmaceutical compositioncomprising:

(i) at least one compound of the present invention,(ii) at least one other active ingredient,as combination products for simultaneous, separate or sequential use.The one or more active ingredients may be such as cited above. Inparticular, the active ingredient may be useful for treatingproliferative diseases such as cancer, psoriasis or fibrosis,advantageously an anti-cancer agent such as an anti-vascular, cytotoxicor anti-angiogenic agent.

The pharmaceutical compositions as described above may be useful fortreating proliferative diseases, such as cancer, psoriasis or fibrosis,particularly cancer.

Finally, the present invention concerns a method for treatingproliferative diseases, such as cancer, psoriasis or fibrosis, andparticularly cancer, comprising the administration, to a patient in needthereof, of a compound of the present invention alone or in combination,advantageously synergistic, with at least one other active ingredient asdefined above.

The present invention also concerns pharmaceutical compositions in theform of nanoparticle formulations. In such embodiments, thepharmaceutical compositions comprise at least one compound of thepresent invention coupled covalently to at least one molecule of ahydrocarbon compound having a squalene structure. In particular, thepharmaceutical compositions comprise at least one compound of thepresent invention coupled covalently to at least one molecule of ahydrocarbon compound having a squalene structure and a polar solvent orwater.

The hydrocarbon compound having a squalene structure and the covalentcoupling of this compound with compounds of the present invention may beas described in WO 2006/090029 or FR 2 924 024.In particular, the hydrocarbon compounds having a squalene structuredesignate linear hydrocarbon structures made up of isoprene units, moreparticularly of six isoprene units. For example, the hydrocarboncompounds having a squalene structure may be squalenic acid andderivatives thereof, particularly substituted derivatives thereof. Thehydrocarbon compounds having a squalene structure may bear a functionalgroup capable of reacting with a functional group present on compoundsof the present invention so as to form a covalent bond between the twocompounds, particularly an ester, ether, thiether, disulphide, phosphateor amide type covalent bond. Alternatively, the covalent bond betweenthe two compounds may be achieved by means of a linker arm. Such linkerarms are well-known to persons skilled in the art.The invention will now be illustrated, in a non-limiting manner, by thefollowing examples.

Examples General Procedure for Synthesizing Compounds I-1 to I-19

4-Chloroquinazoline (1 mmol) is added to a solution of N-tosylhydrazone(1.5 mmol), t-BuOLi (2.2 mmol), PdCl₂(CH₃CN)₂ (0.1 mmol), and dppf (0.2mmol) in dioxane (1 mL). The reaction medium is sealed then heated at90° C. for 2 hours before being returned to room temperature. Thesuspension obtained is filtered on a Celite column (eluent AcOEt) toseparate the inorganic salts. After evaporation of the solvents undervacuum, the residue formed is chromatographed on a silica gel column.

4-[1-(4-Methoxy-phenyl)-vinyl]-2-methyl-quinazoline I-1

Yellow oil, 50%. TLC: Rf 0.3 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1608, 1554, 1511, 1250, 1180. ¹H NMR (300 MHz, CDCl₃): 7.97 (d, 1H,J=8.5 Hz), 7.87 (d, 1H, J=8.5 Hz), 7.81 (td, 1H, J=8.5 Hz, J=0.9 Hz),7.41 (td, 1H, J=8.3 Hz, J=0.9 Hz), 7.23 (d, 2H, J=8.9 Hz), 6.82 (d, 2H,J=8.9 Hz), 6.04 (s, 1H), 5.46 (s, 1H), 3.78 (s, 3H), 2.93 (s, 3H). ¹³CNMR (75 MHz, CDCl₃): 169.9, 164.1, 159.8, 151.1, 145.0, 133.9, 131.4,128.1, 128.0 (2), 127.1, 126.7, 121.7, 116.9, 114.1 (2C), 55.4, 26.8.m/z MS (ESI⁺): 277 (M+H)⁺.

4-[1-(3,4-Dimethoxy-phenyl)-vinyl]-2-methyl-quinazoline I-2

Yellow oil, 48%. TLC: Rf 0.1 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1554, 1514, 1464, 1326, 1221, 1143. ¹H NMR (300 MHz, CDCl₃): 7.96 (d,1H, J=8.4 Hz), 7.87-7.78 (m, 2H), 7.41 (t, 1H, J=7.6 Hz), 6.97 (d, 1H,J=1.6 Hz), 6.76-6.68 (m, 2H), 6.03 (s, 1H), 5.48 (s, 1H), 3.84 (s, 3H),3.41 (s, 3H), 2.93 (s, 3H). ¹³C NMR (75 MHz, CDCl₃): 169.7, 164.1,151.1, 149.5, 149.1, 145.3, 133.9, 131.8, 128.1, 127.0, 126.7, 121.7,120.1, 117.4, 111.1, 109.5, 56.0 (2C), 26.8. m/z MS (APCI⁺): 307 (M+H)⁺.

2-Methyl-4-(1-(4-(trifluoromethoxy)phenyl)vinyl)quinazoline I-3

Yellow oil, 78%. TLC: Rf 0.3 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1554, 1509, 1490, 1254, 1207, 1164. ¹H NMR (300 MHz, CDCl₃): 8.00 (d,1H, J=8.2 Hz), 7.85-7.82 (m, 2H), 7.49-7.43 (td, 1H, J=7.4 Hz, J=1.1Hz), 7.36 (d, 2H, J=8.9 Hz), 7.15 (d, 2H, J=8.1 Hz), 6.16 (s, 1H), 5.62(s, 1H), 2.93 (s, 3H). ¹³C NMR (75 MHz, CDCl₃): 169.2, 164.1, 151.1,149.3, 144.3, 137.4, 134.3, 128.3 (2C), 128.2, 127.1, 126.7, 121.6,121.1 (2C), 120.4 (q, J=256.5 Hz), 119.9, 26.8. ¹⁹F NMR (188 MHz,CDCl₃): −58.23. m/z MS (APCI⁺): 331 (M+H)⁺.

2-Methyl-4-(1-(4-(methylthio)phenyl)vinyl)quinazoline I-4

Yellow oil, 43%. TLC: Rf 0.4 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1751, 1554, 1492, 1326, 1215. ¹H NMR (300 MHz, CDCl₃): 7.96 (d, 1H,J=8.4 Hz), 7.83-7.76 (m, 2H), 7.42-7.36 (m, 1H), 7.20 (d, 2H, J=8.6 Hz),7.14 (d, 2H, J=8.7 Hz), 6.09 (s, 1H), 5.52 (s, 1H), 2.91 (s, 3H), 2.42(s, 3H). ¹³C NMR (75 MHz, CDCl₃): 169.4, 164.1, 151.1, 145.0, 139.2,135.4, 133.9, 128.1, 127.0 (2C), 126.9, 126.8, 126.3 (2C), 121.6, 118.1,26.7, 15.5. m/z MS (APCI⁺): 293 (M+H)⁺.

4-[1-(3-Fluoro-4-methoxy-phenyl)-vinyl]-2-methyl-quinazoline I-5

Yellow oil, 46%. TLC: Rf 0.2 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1615, 1554, 1517, 1275, 1134, 1027. ¹H NMR (300 MHz, CDCl₃): 7.99 (d,1H, J=8.3 Hz), 7.86-7.80 (m, 2H), 7.46-7.41 (td, 1H, J=7.2 Hz, J=1.0Hz), 7.13 (dd, 1H, J=12.5 Hz, J=2.2 Hz), 6.96-6.92 (m, 1H), 6.85 (t, 1H,J=8.5 Hz), 6.06 (s, 1H), 5.51 (s, 1H), 3.86 (s, 3H), 2.93 (s, 3H). ¹³CNMR (75 MHz, CDCl₃): 169.3, 164.1, 154.1, 150.9 (d, 1C, J=23.7 Hz),147.9 (d, 1C, J=11.1 Hz), 144.1, 134.1, 131.9 (d, 1C, J=6.3 Hz), 128.2,126.9, 126.8, 122.9 (d, 1CH, J=3.3 Hz), 121.6, 118.1, 114.3 (d, 1C,J=19.4 Hz), 113.3 (d, 1C, J=1.9 Hz), 56.4, 26.7. ¹⁹F NMR (188 MHz,CDCl₃): −132.7. m/z MS (APCI⁺): 295 (M+H)⁺.

2-Methoxy-5-(1-(2-methylquinazolin-4-yl)vinyl)phenol I-6

4-Chloroquinazoline (1 mmol) is added to a mixture containing silylatedN-tosylhydrazone (1.5 mmol), t-BuOLi (2.2 mmol), PdCl₂(CH₃CN)₂ (0.1mmol) and dppf (0.2 mmol) in dioxane (1 mL). The reaction medium issealed then heated at 90° C. for 2 hours before being returned to roomtemperature. The suspension obtained is filtered on a Celite column(eluent AcOEt) to separate the inorganic salts. After evaporation of thesolvents under vacuum, the residue formed is dissolved in MeOH (1 mL)then K₂CO₃ (2 mmol) is added and the reaction medium is stirred at roomtemperature for 6 hours. The suspension thus formed is filtered, theorganic solvents are evaporated and the residue formed ischromatographed on a silica gel column.Yellow oil, 34%. TLC: Rf 0.1 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1615, 1554, 1512, 1439, 1279, 1135. ¹H NMR (300 MHz, CDCl₃): 7.98 (d,1H, J=8.4 Hz), 7.87 (d, 1H, J=8.3 Hz), 7.81 (td, 1H, J=7.1 Hz, J=1.2Hz), 7.42 (t, 1H, J=7.4 Hz), 6.92 (s, 1H), 6.75 (s, 2H), 6.12 (brs, 1H),6.04 (s, 1H), 5.46 (s, 1H), 3.87 (s, 3H), 2.90 (s, 3H). ¹³C NMR (75 MHz,CDCl₃): 170.1, 164.0, 150.7, 147.1, 145.9, 145.0, 134.1, 132.3, 127.8,127.1, 126.9, 121.8, 118.9, 117.4, 113.0, 110.8, 56.1, 26.5. m/z MS(ESI⁺): 293 (M+H)⁺.

2-Methoxy-5-(1-(2-methylquinazolin-4-yl)vinyl)aniline I-7

4-Chloroquinazoline (1 mmol) is added to a mixture containingN-tosylhydrazone NH-acetyl (1.5 mmol), t-BuOLi (2.2 mmol), PdCl₂(CH₃CN)₂(0.1 mmol) and dppf (0.2 mmol) in dioxane (1 mL). The reaction medium issealed then heated at 90° C. for 2 hours before being returned to roomtemperature. The suspension obtained is filtered on a Celite column(eluent AcOEt) to separate the inorganic salts. After evaporation of thesolvents under vacuum, the residue formed is dissolved in MeOH (1 mL)then KOH (20 mmol) is added and the reaction medium is sealed thenstirred at 100° C. for 12 hours. The suspension thus formed is filtered,the organic solvents are evaporated and the residue formed ischromatographed on a silica gel column.Yellow oil, 27%. TLC: Rf 0.1 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1614, 1567, 1553, 1441, 1330, 1219. ¹H NMR (300 MHz, CD₃COCD₃):7.94-7.84 (m, 3H), 7.52-7.46 (m, 1H), 6.76-6.72 (m, 2H), 6.54 (dd, 1H,J=8.3 Hz, J=2.3 Hz), 5.96 (s, 1H), 5.33 (s, 1H), 4.41 (brs, 2H), 3.81(s, 3H), 2.79 (s, 3H). ¹³C NMR (75 MHz, CD₃COCD₃): 170.7, 164.6, 152.1,148.2, 147.1, 138.5, 134.4, 133.1, 128.9, 127.8, 127.3, 122.9, 116.6,116.1, 113.0, 111.0, 55.8, 26.6. m/z MS (APCI⁺): 292 (M+H)⁺.

2-Methyl-4-(1-(naphthalen-2-yl)vinyl)quinazoline I-8

White solid, 36%. M.p.: 113.6° C. TLC: Rf 0.2 (Cyclohexane/EtOAc, 7/3).IR (neat, cm⁻¹): 1614, 1568, 1490, 1325, 1168, 906. ¹H NMR (300 MHz,CDCl₃): 8.01 (d, 1H, J=8.4 Hz), 7.86 (d, 1H, J=8.4 Hz), 7.83-7.77 (m,3H), 7.68-7.65 (m, 1H), 7.60-7.57 (m, 2H), 7.46-7.34 (m, 3H), 6.28 (s,1H), 5.68 (s, 1H), 2.98 (s, 3H). ¹³C NMR (75 MHz, CDCl₃): 169.6, 164.1,151.1, 145.6, 136.1, 134.0, 133.3, 133.1, 128.5, 128.4, 128.0, 127.6,127.0, 126.8, 126.5, 126.4, 126.4, 124.1, 121.7, 119.3, 26.8. m/z MS(ESI⁺): 297 (M+H)⁺.

2-Methyl-4-(1-(1-methyl-1H-indol-5-yl)vinyl)quinazoline I-9

Reddish oil, 46%. TLC: Rf 0.2 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1612, 1565, 1490, 1332, 1246. ¹H NMR (300 MHz, CDCl₃): 8.00 (d, 1H,J=8.4 Hz), 7.91 (dd, 1H, J=8.4 Hz, J=0.7 Hz), 7.83-7.77 (ddd, 1H, J=8.4Hz, J=6.9 Hz, J=1.4 Hz), 7.50-7.49 (m, 1H), 7.40-7.34 (ddd, 1H, J=8.4Hz, J=6.7 Hz, J=1.3 Hz), 7.30-7.27 (m, 2H), 7.03 (d, 1H, J=3.1 Hz), 6.41(d, 1H, J=3.1 Hz), 6.12 (d, 1H, J=0.7 Hz), 5.53 (d, 1H, J=0.7 Hz), 3.77(s, 3H), 2.99 (s, 3H). ¹³C NMR (75 MHz, CDCl₃): 170.6, 164.1, 151.0,146.8, 136.7, 133.7, 130.7, 129.7, 128.6, 127.9, 127.4, 126.6, 121.9,120.6, 119.7, 116.7, 109.5, 101.7, 33.0, 26.8. m/z MS (APCI⁺): 300(M+H)⁺.

2-Chloro-4-[1-(4-methoxy-phenyl)-vinyl]-quinazoline I-10

Brown oil, 46%. TLC: Rf 0.4 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1673, 1597, 1511, 1246, 1175. ¹H NMR (300 MHz, CDCl₃): 8.0 (dd, 1H,J=8.5 Hz, J=1.1 Hz), 7.90-7.85 (m, 2H), 7.49 (td, 1H, J=8.5 Hz, J=1.1Hz), 7.23 (d, 2H, J=8.8 Hz), 6.84 (d, 2H, J=8.8 Hz), 6.06 (s, 1H), 5.55(s, 1H), 3.79 (s, 3H). ¹³C NMR (75 MHz, CDCl₃): 173.1, 160.1, 157.3,152.7, 144.2, 135.1, 131.1, 128.1 (2C), 128.1, 127.9, 127.5, 122.4,118.6, 114.3 (2C), 55.4. m/z MS (APCI⁺): 297 (M+H)⁺.

2-Chloro-4-[1-(3,4-dimethoxy-phenyl)-vinyl]-quinazoline I-11

Yellow oil, 19%. TLC: Rf 0.3 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1665, 1595, 1563, 1514, 1465, 1264, 1143. ¹H NMR (300 MHz, CDCl₃): 8.00(d, 1H, J=8.9 Hz), 7.91-7.85 (m, 2H), 7.49 (td, 1H, J=8.2 Hz, J=1.2 Hz),6.95 (d, 1H, J=2.0 Hz), 6.77 (d, 1H, J=8.4 Hz), 6.71 (dd, 1H, J=8.3 Hz,J=2.0 Hz), 6.05 (s, 1H), 5.59 (s, 1H), 3.86 (s, 3H), 3.83 (s, 3H). ¹³CNMR (75 MHz, CDCl₃): 172.8, 157.2, 152.7, 149.7, 149.2, 144.5, 135.2,131.5, 128.0, 128.0, 127.4, 122.4, 120.2, 119.2, 111.2, 109.6, 56.1,56.0. m/z MS (APCI⁺): 327 (M+H)⁺.

2-Chloro-4-[1-(3,4,5-trimethoxy-phenyl)-vinyl]-quinazoline I-12

White solid, 34%. M.p.: 135-136° C. TLC: Rf 0.4 (Cyclohexane/EtOAc,7/3). IR (neat, cm⁻¹): 1580, 1528, 1401, 1316, 1277, 1243, 1121. ¹H NMR(300 MHz, CDCl₃): 8.01 (d, 1H, J=8.9 Hz), 7.92-7.87 (m, 2H), 7.55-7.49(m, 1H), 6.52 (s, 2H), 6.09 (s, 1H), 5.66 (s, 1H), 3.85 (s, 3H), 3.66(s, 6H). ¹³C NMR (75 MHz, CDCl₃): 172.4, 157.2, 153.5 (2C), 152.7,144.9, 138.8, 135.3, 134.3, 128.1 (2C), 127.3, 122.3, 120.8, 104.5 (2C),61.1, 56.4 (2C). m/z MS (ESI⁺): 357 (M+H)⁺.

4-[1-(3,5-Dimethoxy-phenyl)-vinyl]-2-methyl-quinazoline I-13

White solid, 36%. F. 89-91° C. Rf 0.3 (Cyclohexane/EtOAc, 7/3). IR(neat, cm⁻¹): 1592, 1460, 1424, 1294, 1206, 1161. ¹H NMR (300 MHz,CDCl₃): 7.96 (d, 1H, J=8.4 Hz), 7.87-7.78 (m, 2H), 7.42 (td, 1H, J=8.1Hz, J=1.0 Hz), 6.45 (d, 2H, J=2.1 Hz), 6.41 (d, 1H, J=2.0 Hz), 6.12 (s,1H), 5.58 (s, 1H), 3.72 (s, 6H), 2.93 (s, 3H). ¹³C NMR (75 MHz, CDCl₃);169.3, 164.1, 161.0 (2C), 151.1, 145.7, 141.0, 134.0, 128.1, 127.0,126.8, 121.7, 119.5, 105.4 (2C), 100.2, 55.5 (2C), 26.8. m/z MS (APCI⁺):307 (M+H)⁺.

2-Methyl-4-[1-(3,4,5-trimethoxy-phenyl)-vinyl]quinazoline I-14

Yellow solid, 40%. F. 127-128° C. Rf 0.2 (Cyclohexane/EtOAc, 7/3). IR(neat, cm⁻¹): 1553, 1506, 1464, 1410, 1331, 1242, 1128. ¹H NMR (300 MHz,CDCl₃): 7.97 (d, 1H, J=8.5 Hz), 7.90-7.80 (m, 2H), 7.45 (td, 1H, J=8.2Hz, J=1.1 Hz), 6.54 (s, 2H), 6.07 (s, 1H), 5.56 (s, 1H), 3.84 (s, 3H),3.72 (s, 6H), 2.94 (s, 3H). ¹³C NMR (75 MHz, CDCl₃): 169.3, 164.1, 153.4(2C), 151.2, 145.7, 138.6, 134.7, 134.0, 128.1, 127.0, 126.9, 121.7,119.0, 104.5 (2C), 61.0, 56.3 (2C), 26.8. m/z MS (ESI⁺): 359 (M+Na)⁺.The compound3-(2-Methoxy-5-(1-(2-methylquinazolin-4-yl)vinyl)phenyl)prop-2-yn-1-olI-15 was prepared in three steps starting with commercial3-iodo-4-methoxyacetophenone (Scheme 1). The first step is a Sonogashirareaction to install the acetylenic functional group. The ketone thusformed is transformed into N-tosylhydrazone derivative then the latteris coupled under palladium-catalysed conditions with 4-chloroquinazolineto yield compound I-15.

Yellow oil, 17%, TLC: Rf 0.1 (Cyclohexane/EtOAc, 1/1). IR (neat, cm⁻¹):3272, 2927, 2840, 2247, 1777, 1599, 1361, 873. ¹H NMR (300 MHz, CDCl₃):8.01 (d, 1H, J=8.3 Hz), 7.82 (m, 2H), 7.46, (t, 1H, J=7.5 Hz), 7.41 (s,1H), 7.22 (d, 1H, J=7.5 Hz), 6.77 (d, 1H, J=8.5 Hz), 6.02 (s, 1H), 5.50(s, 1H), 4.50 (s, 2H), 3.78 (s, 3H), 2.90 (s, 3H), OH not seen). ¹³C NMR(300 MHz, CDCl₃): 169.5, 163.9, 160.0, 150.7, 144.1, 134.1, 131.9,131.2, 128.4, 127.8, 126.9, 126.8, 121.5, 117.9, 112.1, 110.7, 92.0,81.3, 55.9, 51.6, 26.5. HRMS (ESI⁺): m/z calculated for C₂₁H₁₉N₂O₂[M+H]⁺ 331.1447. found 331.1441.The compound 4-(1-(4-Methoxyphenyl)vinyl)quinoline-2-carbonitrile I-16is prepared from 2,4-dichloroquinoline by first carrying out a selectivecyanation reaction at position C₂ (scheme 2). The2-cyano-4-chloroquinoline thus formed with 83% yield is then coupledwith N-tosylhydrazone of 4-methoxyacetophenone.Yellow oil, 62%, TLC: Rf 0.4 (Cyclohexane/EtOAc, 9/1). R (neat, cm⁻¹):3062, 3004, 2958, 2934, 2838, 2236, 1675, 1605, 1576, 1545, 1511, 1459,1218, 1153, 1098, 909. ¹H NMR (300 MHz, CDCl₃): 8.20 (d, 1H, J=9.1 Hz),7.91-7.78 (m, 2H), 7.65 (s, 1H), 7.61-7.50 (m, 1H), 7.18 (d, 2H, J=9.0Hz), 682 (d, 2H, J=9.0 Hz), 6.01 (s, 1H), 5.36 (s, 1H), 3.79 (s, 3H).¹³C NMR (300 MHz, CDCl₃): 160.1, 150.8, 148.7, 144.3, 133.7, 131.7,131.1, 130.4, 129.4, 127.9 (2C), 126.4, 123.8, 117.7, 116.6, 114.3 (2C),55.4. HRMS (ESI⁺): m/z calculated for C₁₉H₁₄N₂O [M+H]⁺ 287.1184. found287.1180.The compound 4-Chloro-2-(1-(4-methoxyphenyl)vinyl)quinoline I-17 isprepared from 2,4-dichloroquinoline)Brown oil, 47%, TLC: Rf 0.5 (Cyclohexane/EtOAc, 9/1). IR (neat, cm⁻¹):3400, 3063, 3001, 2956, 2933, 2907, 2836, 1939, 1652, 1607, 1440, 1245,907, 846. ¹H NMR (300 MHz, CDCl₃): 8.25-8.17 (m, 2H), 7.81-7.75 (td, 1H,J=6.9 Hz, J=1.5 Hz), 7.69-7.61 (td, 1H, J=6.9 Hz, J=1.5 Hz), 7.50 (s,1H), 7.30 (d, 2H, J=9.0 Hz), 6.89 (d, 2H, J=9.0 Hz), 6.04 (d, 1H, J=1.3Hz), 5.75 (d, 1H, J=1.3 Hz), 3.79 (s, 3H). ¹³C NMR (300 MHz, CDCl₃):159.8, 159.1, 148.5, 147.9, 143.0, 132.0, 130.8, 130.0, 129.7 (2C),127.6, 125.7, 124.0, 121.4, 118.7, 114.0 (2C), 55.5. HRMS (ESI⁺): m/zcalculated for C₁₈H₁₅ClNO [M+H]⁺ 296.0842. found 296.0842.The compound 1-(1-(4-Methoxyphenyl)vinyl)-3-methylisoquinoline I-18 isprepared from 1-chloro-3-methylisoquinoline) White solid, 40%, F=66° C.,TLC: Rf 0.42 (Cyclohexane/EtOAc, 3/7). IR (neat, cm⁻¹): 2955, 2835,1621, 1606, 1589, 1464, 1298, 1285, 1098, 879. ¹H NMR (300 MHz, CDCl₃):7.82 (d, 1H, J=9.0 Hz), 7.66 (d, 1H, J=9.0 Hz), 7.51 (t, 1H, J=9.0 Hz),7.40 (s, 1H), 7.27 (t, 1H, J=9.0 Hz), 7.16 (d, 2H, J=9.1 Hz), 6.69 (d,2H, J=9.1 Hz), 5.94 (s, 1H), 5.31 (s, 1H), 3.69 (s, 3H), 2.65 (s, 3H).¹³C NMR (300 MHz, CDCl₃): 160.9, 159.4, 150.9, 146.6, 137.2, 132.5,130.0, 127.8 (2C), 127.4, 126.2, 126.1, 125.4, 118.2, 115.4, 113.7 (2C),55.2, 24.5. HRMS (ESI⁺): m/z calculated for C₁₉H₁₈NO [M+H]⁺ 276.1388.found 276.1385.The compound 4-(1-(4-Methoxyphenyl)vinyl)-2-methylquinoline I-19 isprepared from 4-chloro-2-methylquinoline)White solid, 65%, F=94° C., TLC: Rf 0.24 (Cyclohexane/EtOAc, 8/2). IR(neat, cm⁻¹): 2961, 2929, 1734, 1651, 1607, 1560, 1440, 1409, 1378,1097, 902, 836. ¹H NMR (300 MHz, CDCl₃): 8.04 (d, 1H, J=8.2 Hz), 7.72(d, 1H, J=8.3 Hz), 7.62 (dd, 1H, J=8.35, J=6.92), 7.22 (t, 1H, J=8.3Hz), 7.18 (m, 3H), 6.80 (d, 2H, J=8.0 Hz), 5.80 (s, 1H), 5.31 (s, 1H),3.77 (s, 3H), 2.76 (s, 3H). ¹³C NMR (300 MHz, CDCl₃): 158.5, 157.7,147.7, 147.1, 144.5, 131.3, 128.2, 127.8, 126.7 (2C), 125.0, 124.5,124.4, 121.3, 113.9, 112.8 (2C), 54.2, 24.3. HRMS (ESI⁺): m/z calculatedfor C₁₉H₁₈NO [M+H]⁺ 276.1388. found 276.1388.

General Procedure for Synthesizing Compounds II-1 to II-4

A solution of 1-quinazoline-1-arylethylenes (I) (1 mmol) in EtOAc (1 mL)is placed under atmospheric pressure of hydrogen in the presence of Pd/C(20% by mass) then stirred for 2 to 5 hours. After filtration on Celite,the solvents are evaporated under vacuum, and the residue formed ischromatographed on a silica gel column.

4-[1-(4-Methoxy-phenyl)-ethyl]-2-methyl-quinazoline II-1

Brown oil, 99%. TLC: Rf 0.4 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):2925, 1612, 1564, 1511, 1463, 1247, 1178. ¹H NMR (300 MHz, CDCl₃): 8.07(d, 1H, J=8.4 Hz), 7.91 (d, 1H, J=8.4 Hz), 7.75-7.69 (m, 1H), 7.45-7.39(m, 1H), 7.24 (d, 2H, J=8.7 Hz), 6.79 (d, 2H, J=8.7 Hz), 4.95 (q, 1H,J=6.9 Hz), 3.71 (s, 3H), 2.92 (s, 3H), 1.78 (d, 3H, J=6.9 Hz). ¹³C NMR(75 MHz, CDCl₃): 172.4, 163.8, 158.2, 150.7, 136.5, 133.0, 128.6 (2C),128.5, 126.4, 124.6, 121.4, 114.1 (2C), 55.2, 42.2, 26.8, 21.3. m/z MS(APCI⁺): 279 (M+H)⁺.

4-[1-(3,4-Dimethoxy-phenyl)-ethyl]-2-methyl-quinazoline II-2

Yellow oil, 97%. TLC: Rf 0.2 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹):1563, 1514, 1491, 1418, 1328, 1262, 1140. ¹H NMR (300 MHz, CDCl₃): 8.10(d, 1H, J=8.2 Hz), 7.90 (d, 1H, J=8.3 Hz), 7.75 (ddd, 1H, J=8.2 Hz,J=6.9 Hz, J=1.2 Hz), 7.45 (ddd, 1H, J=8.2 Hz, J=6.8 Hz, J=1.1 Hz), 6.92(d, 1H, J=2.0 Hz), 6.86 (dd, 1H, J=8.2 Hz, J=2.0 Hz), 6.75 (d, 1H, J=8.2Hz), 4.94 (q, 1H, J=6.9 Hz), 3.82 (s, 3H), 3.81 (s, 3H), 2.91 (s, 3H),1.78 (d, 3H, J=6.9 Hz). ¹³C NMR (75 MHz, CDCl₃): 172.3, 163.8, 150.8,149.1, 147.8, 137.0, 133.2, 128.6, 126.5, 124.7, 121.4, 119.8, 111.3,111.0, 56.0, 55.9, 42.6, 26.9, 21.4. m/z MS (APCI⁺): 309 (M+H)⁺.

2-Methyl-4-[1-(3,4,5-trimethoxy-phenyl)-ethyl]-quinazoline II-3

Yellow oil, 97%. Rf 0.2 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹): 1587,1564, 1492, 1461, 1420, 1328, 1238, 1122. ¹H NMR (300 MHz, CDCl₃): 8.1(d, 1H, J=8.2 Hz), 7.91 (d, 1H, J=8.3 Hz), 7.76 (ddd, 1H, J=8.2 Hz,J=7.1 Hz, J=1.3 Hz), 7.47 (ddd, 1H, J=8.2 Hz, J=7.0 Hz, J=1.2 Hz), 6.58(s, 2H), 4.91 (q, 1H, J=6.9 Hz), 3.79 (s, 6H), 3.77 (s, 3H), 2.91 (s,3H), 1.78 (d, 3H, J=6.9 Hz). ¹³C NMR (75 MHz, CDCl₃): 172.0, 163.8,153.3 (2C), 150.8, 140.0, 136.8, 133.2, 128.6, 126.6, 124.6, 121.5,105.0 (2C), 60.9, 56.2 (2C), 43.2, 26.8, 21.5. m/z MS (APCI⁺): 339(M+H)⁺.

4-[1-(3,5-Dimethoxy-phenyl)-ethyl]-2-methyl-quinazoline II-4

Brown oil, 89%. Rf 0.3 (Cyclohexane/EtOAc, 7/3). IR (neat, cm⁻¹): 1609,1593, 1564, 1428, 1205, 1157. ¹H NMR (300 MHz, CDCl₃): 8.08 (d, 1H,J=8.2 Hz), 7.90 (d, 1H, J=8.4 Hz), 7.77-7.72 (ddd, 1H, J=8.2 Hz, J=6.9Hz, J=1.3 Hz), 7.44 (ddd, 1H, J=8.3 Hz, J=7.0 Hz, J=1.0 Hz), 6.49 (d,2H, J=2.2 Hz), 6.27 (t, 1H, J=2.2 Hz), 4.90 (q, 1H, J=6.9 Hz), 3.73 (s,6H), 2.91 (s, 3H), 1.78 (d, 3H, J=6.9 Hz). ¹³C NMR (75 MHz, CDCl₃):171.8, 163.8, 161.0 (2C), 150.7, 146.9, 133.2, 128.5, 126.6, 124.7,121.6, 106.2 (2C), 98.0, 55.4 (2C), 43.4, 26.9, 21.1. m/z MS (APCI⁺):309 (M+H)⁺.

Cytotoxicity Protocol.

U87, H1299 and MDA-MB231 cells are grown in Dulbecco minimal essentialmedium (DMEM) containing 4.5 g/L glucose and supplemented with 10%foetal calf serum and 1% glutamine. K562 and HCT116 cells are grown inRPMI 1640 medium containing 10% foetal calf serum and 1% glutamine. Allthe cell lines are maintained in culture at 37° C. in a humid atmospherecontaining 5% CO₂. The cells are inoculated in 96-well culture plates inan amount of 5000 cells per well in 50 μL of culture medium. After 24hours of culture, the molecule to be tested, dissolved in DMSO anddiluted 1/1000 in suitable culture medium, is added in each well in anamount of 50 μL per well. After 72 hours of incubation, 20 μL ofresazurin is added in each well. After 1 hour of incubation, emittedfluorescence is measured at 590 nm after excitation at 560 nm using theVictor fluorescence reader (Perkin-Elmer, USA). Each concentration istested three times and each experiment is repeated three times.

Protocol for Inhibition of Polymerisation of Tubulin.

When tubulin is placed at a temperature of 0° C., it is in the form offree dimers. On the other hand, when the temperature is suddenlyincreased to 37° C., the dimers assemble into microtubules. The IPT ofthe various synthesized analogues was determined on a soluble tubulinaccording to the method of Shelanski⁷ from sheep brains where itconstitutes 20 to 25% of soluble proteins. The “antitubulin” activity isevaluated according to the method of Gaskin⁸ by measuring turbidity,which is proportional to the microtubule concentration in thesuspension. Turbidity is measured by absorption of the suspension at 350nm using a spectrophotometer fitted with a cellthermostatically-controlled at 37° C. The various samples were dissolvedin DMSO and incubated 10 minutes at 37° C. then 5 minutes at 0° C.The tubulin assembly and disassembly curves are obtained by absorptionkinetics at 350 nm as a function of time. The IC₅₀ determinedgraphically is defined as the concentration of inhibitor necessary todecrease by 50% the maximum rate of assembly of the control tubulin.

In Vitro Biological Study of Compounds of the Present Invention

Their effects on the proliferation of various human cancer cell linesand their capacity to inhibit the polymerisation of tubulin were tested.

A. Cytotoxic Activity

The cytotoxic activity of the compounds was studied in vitro principallyon the HCT116 human cancer cell line (colorectal cancer). The lineselected for this study was incubated at 37° C. in the presence of oneof the compounds added in the culture medium at various concentrations.The inhibitory concentration inducing the death of 50% of the cells(IC₅₀) was determined after 72 hours of incubation (Table 1) for eachcompound. The set of experiments carried out made it possible todetermine the degree of toxicity of the compounds tested by takingisoCA-4 as the reference. In view of the results, it appears thatseveral compounds have IC₅₀ values at a nanomolar level and thatcompounds I-5, I-6, I-7, I-15, I-16, I-19 are the most cytotoxic withIC₅₀ values below 20 nM. It should be noted that the firststructure-activity relationships are in perfect agreement with theprevious results observed in the isoCA-4 and isoerianin series, namelythat the most effective B rings are substituted at C-4 with a OMe groupand at C-3 by a fluorine atom (I-5), a OH (I-6) or NH₂ (I-7) group.Moreover, by comparing the results obtained with the cytotoxic compoundsI-1 and I-10, it appears that the quinazoline nucleus may be substitutedat C-2 with a methyl group or a chlorine atom with no loss of efficacy(I-1 and I-10: IC₅₀=40 nM). Lastly, when these first results areexamined, it appears that the reduction of the double bond locatedbetween the two aromatic nuclei yields “reduced” type II derivativesabout 10 times less cytotoxic than their precursors I as we had alreadynoted during the reduction of isoCA-4 derivatives to isoerianin-typederivatives. Molecule I-15 shows that the introduction of an acetyleniccarbon-containing substituent at C3′ considerably improves the cytotoxicactivity compared to I-1. In addition, replacing the quinazoline nucleiwith other heterocycles made it possible to identify moleculesexhibiting better cytotoxic activities at sub-nanomolar concentrations.

TABLE 1 Cytotoxicity results of the compounds on the HCT116 human cancercell line (colorectal cancer). Cytotoxic activity (IC₅₀) is expressed innMol/L and corresponds to the concentration of the compounds whichinduces 50% cell death after 72 hours of incubation. HCT116 CompoundsIC₅₀ (nM)

I-1  38

I-2  460

I-3 2300

I-4  52

I-5  18

I-6  10

I-7  10

I-8  130

I-9  27

I-10  34

I-11  700

I-12  900

II.1  250

II-2  950

I-15 1, 5

I-16 1, 6

I-17  95

I-18  87

I-19   ≦1 isoCA-4   2

Compounds I-1 and I-10 bearing a methyl substituent and a chlorine atomat C-2 and which had equivalent levels of cytotoxicity (IC₅₀=40 nM) withrespect to the HCT116 cell line were also evaluated on a K562 chronicmyeloid leukaemia line and a H1299 non-small-cell lung cancer line.Compounds I-15, I-16 and I-19 were also evaluated on a K562 chronicmyeloid leukaemia line and a K562R imatinib-resistant chronic myeloidleukaemia line. The first cytotoxicity results are reported in Table 2below.

TABLE 2 Cytotoxicity results of compounds I-1 and I-10 on three humancancer cell lines HCT116 (colorectal cancer), K562 (chronic myeloidleukaemia) and H1299 (non-small-cell lung cancer). Cytotoxic activity(IC₅₀) is expressed in nMol/L and corresponds to the concentration ofthe compounds which induces 50% cell death after 72 hours of incubation.HCT116 IC₅₀ K562 IC₅₀ K562R IC₅₀ H1299 IC₅₀ Compounds (nM) (nM) (nM)(nM) I-1 38 34 n/a 22 I-10 34 38 n/a 25 I-15 1.5 8 ≦1 n/a I-16 1.6 9 ≦1n/a I-19 ≦1 17 0.7 n/a isoCA-4 2 4  5The results indicate that the cytotoxicity of I-1 and I-10 with respectto the K562 and H1299 lines is similar to that observed with the HCT116line (IC₅₀ between 22 and 38 nM).Replacing the quinazoline nucleus (I-1) with a quinoline nucleus yieldedcompound 1-19 having sub-nanomolar cytotoxic activities. It should benoted that replacing the C2 methyl group of compound 1-19 with a cyanogroup (1-16) has no effect on cytotoxic activity.

B. Inhibition of Polymerisation of Tubulin

The most cytotoxic compounds were selected for tests of inhibition ofpolymerisation of tubulin. These tests were carried out under thedirection of Dr J. Dubois at Gif-sur-Yvette, France. Tubulin is purifiedfrom ewe brains according to the method of Shelanski⁹ by twoassembly-disassembly cycles. The stock solution (15-20 mg/mL), stored at−196° C., is thawed and diluted in assembly buffer (0.1 M MES, 0.5 mMMgCl₂, 1 mM EGTA, and 1 mM GTP, pH 6.6) to have a final concentration of10 μM. Tubulin assembly is monitored by fluorescence on 96-well platesaccording to the method of Barron et al.¹⁰ Inhibitor (DMSO, 1 μL) isadded to the tubulin solution (10 μM, 100 μL per well) and the solutionis incubated 45 minutes at room temperature. GTP (1 mM final) is thenadded, the solution is mixed rapidly and fluorescence (λ_(ex)=350 nm,λ_(em)=440 nm) is measured on a Wallac Victor fluorometer (PerkinElmer). The inhibition of 50% of the maximum rate of assembly (IC₅₀) isdetermined in duplicate or triplicate on 10 concentrations bracketingthe IC₅₀. The results are recorded in Table 3.

TABLE 3 Results of the inhibition of the polymerisation of tubulin (IPT)by the selected compounds I Compounds IPT (IC₅₀) μM

I-1 1.8

I-4 2.9

I-5 1.0

I-6 0.6

I-7 2.3

I-8 2.2

I-9 1.6

I-10 1.9

I-15 1.2

I-16 0.96

I-19 1.09 isoCA-4 1.0 CA-4 1.0

It may be noted that all of the selected compounds inhibit thepolymerisation of tubulin at micromolar concentrations (0.6 μM<IC₅₀<2.9μM). In particular, compound I-6, which proved to be very cytotoxic withrespect to the HCT116 line, inhibits the assembly of tubulin with asub-micromolar IC₅₀ (IC₅₀=0.6 μM).

Molecule I-15 shows that the introduction of an acetyleniccarbon-containing substituent at C3′ considerably improves the ITPactivity compared to I-1. In addition, replacing the quinazoline nucleuswith other heterocycles made it possible to identify moleculesexhibiting a capacity to inhibit the polymerisation of tubulin atsub-micromolar concentrations.

REFERENCES

-   1. G. R. Pettit et al J. Nat. Prod. 1987, 50, 119.-   2. Mc Gown et al Cancer Chemother. Pharmacol. 1990, 26, 79.-   3. G. G. Dark et al Cancer Res. 1997, 57, 1829.-   4. (a) S. Messaoudi et al J. Med. Chem. 2009, 52, 4538. (b) M. Alami    et al WO 2008/122620 (c) A. Hamze et al ChemMedChem. 2009, 4, 1912.-   5. M. Soussi et al ChemMedChem, 2011, 6, 1781.-   6. M. Alami et al WO 2009/147217.-   7. Shelanski, M. C.; Gaskin, F.; Cantor, C. R. Proc. Natl. Acad.    Sci. USA, 1973, 70, 765-768.-   8. Gaskin, F.; Cantor, C. R.; Shelanski, M. C. J. Biol. Mol., 1974,    89, 737-755.-   9. Shelanski, M. L.; Gaskin, F.; Cantor, C. R. Proc. Natl. Acad.    Sci. USA, 1973, 70, 765-768.-   10. Barron, D. M.; Chatterjee, S. K.; Ravindra, R.; Roof, R.;    Baloglu, E.; Kingston, D. G. I.; Bane, S. Anal. Biochem. 2003, 315    49-56.

1.-16. (canceled)
 17. A compound of formula (I):

wherein: R₂ and R₃ are different, and one of R₂ and R₃ is a group A₁ of the following formula:

wherein: the dashed bond is present or absent; Z₁ and Z₂ are, independently of each other, a hydrogen atom, a halogen atom or a methyl group; (Het)Aryl is an aryl or heteroaryl group, said heteroaryl being selected from indolyl, benzothiophenyl and benzofuranyl groups, wherein said aryl or heteroaryl may be substituted with one or more substituents selected from a halogen atom, a hydroxyl group, —OMe, —SMe, —OCX₃ with X indicating a halogen atom, —NH₂,

with R indicating a C₁-C₆ alkyl group,

wherein m is 1 or 2,

wherein m is 1 or 2, and

and the other of R₂ and R₃ is: a hydrogen atom; a halogen atom; a hydroxyl group; a cyano group; a —COYR′ group wherein Y is O or N and R′ is H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, or a C₂-C₄ alkynyl group; a —SO₂NR′R″ group wherein R′, R″ each is, independently of each other, H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, or a C₂-C₄ alkynyl group; a —NHSO₂R′ group wherein R′ is a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group, an aryl group, or a heteroaryl group; a C₁-C₆ alkyl group; a C₂-C₄ alkenyl group; a C₂-C₄ alkynyl group; a C₁-C₆ alkoxy group; or a —NR′R″ group wherein R′ and R″ are, independently of each other, a hydrogen or an alkyl group; A and E are each a hydrogen atom, or A and E take part in the structure of a fused aromatic ring of the following formula:

wherein A, B and E are, independently of each other, a carbon or nitrogen atom, n is 0 or 1, and R₄, R₅, R₆, R₇ are as described below; X, Y and Z are, independently of each other, a carbon or nitrogen atom provided that if X and Z is a nitrogen atom, Y is a carbon atom; at least one of A, B, E, X, Y and Z is a nitrogen atom; R₁, R₄, R₅, R₆, R₇, if present, are, independently of each other: a hydrogen atom; a halogen atom; a hydroxyl group; a C₁-C₆ alkyl group; a C₂-C₄ alkenyl group; a C₂-C₄ alkynyl group; a C₁-C₆ alkoxy group; or a —NR′R″ group wherein R′ and R″ are, independently of each other, a hydrogen or a C₁-C₆ alkyl group; as well as pharmaceutically acceptable salts, stereoisomers and prodrugs thereof.
 18. The compound according to claim 17, wherein the dashed bond is present.
 19. The compound according to claim 17 having the following formula (I′):


20. The compound according to claim 19 having the following formula (Ib):

wherein: the dashed bond is present or absent; R₂ is: a hydrogen atom; a halogen atom; a hydroxyl group; a cyano group; a —COYR′ group wherein Y is O or N, and R′ is H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, or a C₂-C₄ alkynyl group; a —SO₂NR′R″ group wherein R′, R″ are each, independently of each other, H or a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, or a C₂-C₄ alkynyl group; a —NHSO₂R′ group wherein R′ is a C₁-C₆ alkyl group, a C₂-C₄ alkenyl group, a C₂-C₄ alkynyl group, an aryl group, or a heteroaryl group; a C₁-C₆ alkyl group; a C₂-C₄ alkenyl group; a C₂-C₄ alkynyl group; a C₁-C₆ alkoxy group; or a —NR′R″ group wherein R′ and R″ are, independently of each other, a hydrogen or an alkyl group; Z₁ and Z₂ are, independently of each other, a hydrogen atom, a halogen atom or a methyl group; (Het)Aryl is an aryl or heteroaryl group, said heteroaryl being selected from indolyl, benzothiophenyl and benzofuranyl groups, wherein said aryl or heteroaryl may be substituted with one or more substituents selected from a halogen atom, a hydroxyl group, —OMe, —SMe, —OCX₃ wherein X is a halogen atom, —NH₂,

wherein R is a C₁-C₆ alkyl group,

wherein m is 1 or 2,

wherein m is 1 or 2 or


21. The compound according to claim 20, wherein the dashed bond is present.
 22. The compound according to claim 17, wherein (Het)Aryl is a phenyl group which may be substituted with one or more substituents selected from a halogen atom, a hydroxyl group, —OMe, —SMe, —OCX₃ wherein X is a halogen atom, —NH₂,

wherein R is a C₁-C₆ alkyl group,

wherein m is 1 or 2,

wherein m is 1 or 2 or


23. The compound according to claim 22, wherein the phenyl group has the following formula:

wherein: R₈ is a hydrogen atom, a halogen atom, a hydroxyl, —OMe, —SMe, —NH₂,

wherein R is a C₁-C₆ alkyl group,

wherein m is 1 or 2,

wherein m is 1 or 2 or

and R₉ is a C₁-C₆ alkoxy group, —SMe or —OCX₃ wherein X is a halogen atom.
 24. The compound according to claim 17 wherein Z₁ and Z₂ are a hydrogen atom.
 25. The compound according to claim 17 wherein Z₁ and Z₂ are a fluorine atom.
 26. The compound according to claim 17 wherein n is 1, X and Z are a nitrogen atom, A and E are, independently of each other, a nitrogen or carbon atom, and Y and B are a carbon atom.
 27. The compound according to claim 17, wherein the compound is selected from:


28. A method for treating or preventing a proliferative disease, comprising administering to a person in need thereof an effective amount of a compound according to claim
 17. 29. The method according to claim 28, wherein the proliferative disease is cancer, psoriasis or fibrosis.
 30. A method for treating or preventing a vascular disease, comprising administering to a person in need thereof an effective amount of a compound according to claim
 17. 31. A conjugate comprising a compound according to claim 17 bound covalently to an antibody.
 32. A pharmaceutical composition comprising a compound according to claim
 17. 33. A pharmaceutical composition comprising a conjugate according to claim
 31. 34. The pharmaceutical composition according to claim 32 further comprising at least one other active ingredient.
 35. The pharmaceutical composition according to claim 34 wherein the other active ingredient is selected from 6-mercaptopurine, fludarabine, cladribine, pentostatin, cytarabine, 5-fluorouracil, gemcitabine, methotrexate, raltitrexed, irinotecan, topotecan, etoposide, daunorubicin, doxorubicin, epirubicin, idarubicin, pirarubicin, mitoxantrone, chlormethine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, busulfan, carmustine, fotemustine, streptozocin, carboplatin, cisplatin, oxaliplatin, procarbazine, dacarbazine, bleomycin, vinblastine, vincristine, vindesine, vinorelbine, paclitaxel, docetaxel, L-asparaginase, flutamide, nilutamide, bicalutamide, cyproterone acetate, triptorelin, leuprorelin, goserelin, buserelin, formestane, aminoglutethimide, anastrazole, letrozole, tamoxifen, octreotide and lanreotide.
 36. A pharmaceutical composition comprising: (i) at least one compound of formula (I) according to claim 17 and (ii) at least one other active ingredient, as a combination product for simultaneous, separate or sequential use.
 37. A method for preparing a compound of formula (I) comprising the following successive steps: a) reaction of a compound of formula (II)

with tosylhydrazine to yield the tosylhydrazone compound of formula (III)

b) coupling of the compound of formula (III) with the compound of formula (IV), (V), (VI) or (VII)

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, X, Y, Z, A, B, E and n are as defined in claim 17 and X₁ is a halogen atom or trifluoromethanesulfonate; c) separation from the reaction medium of compound (I) obtained following the previous step.
 38. The method according to claim 37, further comprising a step of reduction after step b) and before step c) to obtain a compound of formula (I) wherein the dashed bond is absent. 